Coupled transcription and translation in eukaryotic cell-free extract

Abstract

A method for producing protein from DNA in a static reaction in which DNA, ribonucleotide triphosphates, magnesium and a RNA polymerase are mixed and incubated during which RNA is transcribed from the DNA and the resulting solution is added to a eukaryotic cell-free extract containing magnesium so that the RNA produced during the incubating step translates into protein and so that further transcription of the DNA is coupled to translation of the resulting RNA into protein.

What is claimed is: 
     
       1. A method for enhanced protein production from a DNA template comprising a first step in which DNA, magnesium, ribonucleotide triphosphates and an RNA polymerase are mixed to form a solution and allowed to incubate for 2-30 minutes during which time RNA is transcribed from the DNA, and   a step following said first step in which said solution is mixed with a eukaryotic cell-free extract containing magnesium, so that further transcription is coupled to simultaneous translation of the RNA into protein.   
     
     
       2. A method for protein production comprising a first step is which DNA template, magnesium, ribonucleotide triphosphates and an RNA polymerase are mixed to form a solution and incubated for 2-30 minutes and   followed by a second step in which said solution is mixed with a eukaryotic cell-free extract containing magnesium sufficient such that RNA is transcribed from said template DNA and the RNA translates into said protein.   
     
     
       3. A method for enhanced protein production as set forth in claims 1 or 2 wherein said eukaryotic cell-free extract is rabbit reticulocyte lysate. 
     
     
       4. A method for enhanced protein production as set forth in claims 1 or 2 wherein said eukaryotic cell-free extract is wheat germ extract.